Importance of porins for biocide efficacy against Mycobacterium smegmatis
Elrike Frenzel1, Stefan Schmidt, Michael Niederweis
1Schülke & Mayr GmbH, Research & Development, 22851 Norderstedt, Germany.
Abstract:
Mycobacteria are among the microorganisms least susceptible to biocides but cause devastating diseases, such as tuberculosis, and increasingly opportunistic infections. The exceptional resistance of mycobacteria to toxic solutes is due to an unusual outer membrane, which acts as an efficient permeability barrier, in synergy with other resistance mechanisms. Porins are channel-forming proteins in the outer membrane of mycobacteria. In this study we used the alamarBlue assay to show that the deletion of Msp porins in isogenic mutants increased the resistance of Mycobacterium smegmatis to isothiazolinones (methylchloroisothiazolinone [MCI]/methylisothiazolinone [MI] and octylisothiazolinone [2-n-octyl-4-isothiazolin-3-one; OIT]), formaldehyde-releasing biocides {hexahydrotriazine [1,3,5-tris (2-hydroxyethyl)-hexahydrotriazine; HHT] and methylenbisoxazolidine [N,N'-methylene-bis-5-(methyloxazolidine); MBO]}, and the lipophilic biocides polyhexamethylene biguanide and octenidine dihydrochloride 2- to 16-fold. Furthermore, the susceptibility of the porin triple mutant against a complex disinfectant was decreased 8-fold compared to wild-type (wt) M. smegmatis. Efficacy testing in the quantitative suspension test EN 14348 revealed 100-fold improved survival of the porin mutant in the presence of this biocide. These findings underline the importance of porins for the susceptibility of M. smegmatis to biocides.
Insights
Mycobacterium outer membrane porins significantly enhance resistance to various biocides. Deleting these porins increases resistance to disinfectants, crucial for understanding mycobacterial survival strategies.
Area of Science:
- Microbiology
- Biochemistry
Background:
- Mycobacteria exhibit exceptional resistance to biocides due to their unique outer membrane barrier.
- Porins, channel-forming proteins in the mycobacterial outer membrane, are implicated in biocide susceptibility.
Purpose of the Study:
- To investigate the role of Msp porins in the biocide resistance of Mycobacterium smegmatis.
- To determine how porin deletion affects susceptibility to a range of biocidal agents.
Main Methods:
- Utilized the alamarBlue assay to quantify biocide resistance in isogenic Mycobacterium smegmatis mutants.
- Employed quantitative suspension tests (EN 14348) to assess disinfectant efficacy against porin mutants.
Main Results:
- Deletion of Msp porins increased Mycobacterium smegmatis resistance to isothiazolinones, formaldehyde-releasers, and lipophilic biocides by 2- to 16-fold.
- A porin triple mutant showed an 8-fold decreased susceptibility to a complex disinfectant, with 100-fold improved survival in suspension tests.
Conclusions:
- Porins play a critical role in determining the susceptibility of Mycobacterium smegmatis to biocides.
- Targeting porins could be a strategy to enhance the efficacy of disinfectants against mycobacteria.
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